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OBJECTIVE The aim of this study was to investigate the bacterial bioburden in experimental in vitro wounds during the application of conventional negative pressure wound therapy (NPWT), with and without antimicrobial dressings (polyhexanide, silver), against NPWT instillation of octenidine. METHOD Experimental wounds produced in an in vitro porcine wound model were homogenously contaminated with bacterial suspension and treated with NPWT and different options. Group A: non-antimicrobial polyurethane foam dressing; group B: antimicrobial polyurethane foam dressing containing silver; group C: antimicrobial gauze dressing containing polyhexanide; group D: non-antimicrobial polyurethane foam dressing intermittently irrigated with octenidine; group E: negative control (non-antimicrobial polyurethane foam dressing without NPWT). Standard biopsies were harvested after 24 and 28 hours. RESULTS This study demonstrated that the use of NPWT with intermitted instillation of octenidine (group D) or application of silver-based polyurethane foam dressings (group B) is significantly superior against Staphylococcus aureus colonisation in experimental wounds compared with non-antimicrobial polyurethane foam dressing (group A) after 48 hours. Surprisingly, the polyhexanide-based dressing (group C) used in this model showed no statistical significant effect compared with the control group (group E) after 24 or 48 hours of treatment. CONCLUSION Both intermitted instillation of octenidine and silver-based dressings in standard NPWT were significantly superior compared with non-antimicrobial polyurethane foam dressings or PHMB coated gauze dressing after 48 hours.